Purification and characterization of a group of five novel peptide serine protease inhibitors from ovaries of the desert locust, Schistocerca gregaria.
The ovary of the desert locust, Schistocerca gregaria, contains multiple inhibitors of serine proteases. Five serine protease inhibitors, designated SGPI-1-5 (Schistocerca gregaria protease inhibitors) were purified from methanolic extracts of mature ovaries and analyzed by mass spectrometry and amino acid sequencing. The revealed primary structures display amino acid similarities and are related to the serine protease inhibitors identified in the hemolymph of Locusta migratoria. All inhibitors show an in vitro inhibiting activity towards alpha-chymotrypsin. In addition, SGPI-1 displays in vitro inhibiting activity towards trypsin, and SGPI-2 is a potent pancreatic elastase inhibitor. Differences in inhibitory specificities towards the locust endogenous serine proteases can be readily attributed to the amino acid sequence within the active region and also to amino acid residues beyond the P1-P'1 bond. A difference in one or two amino acid residues around the reactive sites results in considerable alteration of the inhibitory specificity. The temporal and spatial distribution of SGPI-1-5 was studied by RP-HPLC analysis. All inhibitors occur in hemolymph, ovaries, testes and fat body of adults but are absent in the gut. They are also present in larval hemolymph and fat body. An antibody raised against SGPI-2 shows positive immunostaining in the ovarian follicle cells.
Hamdaoui
A
A
Faculty of Science Semlalia, Université Cadi Ayyad, Marrakech, Morocco.
Wataleb
S
S
Devreese
B
B
Chiou
S J
SJ
Vanden Broeck
J
J
Van Beeumen
J
J
De Loof
A
A
Schoofs
L
L
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D005260:Female; D006110:Grasshoppers; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000302:isolation & purification / Q000494:pharmacology; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010053:Ovary / Q000737:chemistry*; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology; D017421:Sequence Analysis; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000302:isolation & purification*; D014361:Trypsin Inhibitors / Q000494:pharmacology
D007527:Isoenzymes; D010455:Peptides; D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors; D002918:Chymotrypsin; D010196:Pancreatic Elastase